Compositions and methods for treating hair loss and delaying aging of skin

ABSTRACT

The invention provides methods of hair loss or inducing new hair growth, as well as methods of preventing hair loss, comprising applying an effective amount of a dihydromyricetin compound to the scalp. The invention also provides methods of delaying or reversing signs of aging skin comprising applying an effective amount of a dihydromyricetin compound to the skin. Also provided are cosmetic products (e.g., skin and hair products) comprising a dihydromyricetin compound and methods of making the same.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication No. 61/671,624, filed Jul. 13, 2012, the disclosure of whichis hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

This invention relates to compositions and methods to treat hair loss,promote hair growth, or delay or reverse the signs of aging skin, andmethods of making related products.

BACKGROUND OF THE INVENTION

Hair loss is a common problem that affects men and women of all agesthroughout the world. One large-scale study in Maryborough, Victoria,Australia, showed the prevalence of mid-frontal hair loss increases withage and affects 73.5% of men and 57% of women aged 80 and over (Gan etal. (2005) “Prevalence of male and female pattern hair loss inMaryborough.” J Investig Dermatol Symp Proc. 10:184-189). According toMedem Medical Library's website, male pattern hair loss or baldness(MPB) affects about 40 million men in the United States. Approximately25% of men begin balding by age 30; two-thirds begin balding by age 60.

Hair growth typically follows a continuously repeating cycle related torest, shedding, and regrowth. However, a variety of factors, includinghormonal changes, poor nutrition, illness, medications, and stress, candisrupt this cycle and lead to excessive hair loss, thinning hair, orbaldness. For example, the hormone dihydrotestosterone or DHT is widelyaccepted to be the main cause of male pattern baldness. Thyroid disease,diabetes, and lupus frequently cause general hair loss. Stress canaggravate alopecia areata, which is characterized by the sudden loss ofhair in round or oval patches about the size of a quarter. A second formstress-induced hair loss is telogen effluvium (TE), which occurs when asudden or severe stress causes an acute increase in the shedding ofhair.

Currently available hair loss treatments have been shown to bemoderately successful in renewing hair growth. However, medications forthe treatment of hair loss, such as minoxidil, procaine, andfinasteride, can comprise pharmaceutical agents that cause undesirableside effects, including irritation, severe allergic reactions, and anincreased risk of prostate cancer in men. Finasteride also posessignificant danger to women of childbearing age and should not behandled by pregnant women. Moreover, surgical procedures can beexpensive, painful, and can carry the risk of complications, such asinfection, scarring, or unnatural appearance of the direction of hairgrowth.

What is needed is a safe, effective, natural treatment that can beadministered topically to prevent and/or reverse hair loss or induce newhair growth.

All references cited herein, including patent applications andpublications, are hereby incorporated by reference in their entirety.

BRIEF SUMMARY OF THE INVENTION

In one aspect, the invention provides a method for treating hair loss orinducing hair growth (such as new hair growth) comprising applying aneffective amount of a dihydromyricetin compound to scalp of anindividual, wherein the dihydromyricetin compound is of formula (I):

wherein each X is independently selected from hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, halo, alkoxyl,heteroalkyl, substituted heteroalkyl, heteroalkenyl, or substitutedheteroalkenyl, or a salt thereof. In some embodiments, the X is hydrogen(i.e., the dihydromyricetin compound is dihydromyricetin). In someembodiments, the X is independently selected from the group consistingof —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —CH(CH₃)₂, —CH₂CH₂CH₂CH₃, —CH₂CH(CH₃)₂,—C(CH₃)₃, —CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₃, —C(CH₃)₂CH₂CH₃,—CH₂CH₂CH(CH₃)₂, —CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃,—CH₂CH₂C(CH₃)₂CH₃, —CH₂CH₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃,—CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₂CH₃, and—CH₂CH₂CH₂CH₂OCH₂CH₃. In some embodiments, the dihydromyricetin compoundis of formula (II):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is of formula (III):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is provided in a hairproduct (e.g., a shampoo, a conditioner, or a hair spray, etc.). In someembodiments, the dihydromyricetin compound in the hair product is in aconcentration of at least about 5% (w/v-%) or at least about 5% (w/w-%).In some embodiments, the dihydromyricetin compound in the hair productis in a concentration of at least about 10% (w/v-%) or at least about10% (w/w-%).

In another aspect, the invention provides a method for preventing hairloss comprising applying an effective amount of a dihydromyricetincompound to scalp of an individual, wherein the dihydromyricetincompound is of formula (I):

wherein X is hydrogen, alkyl, substituted alkyl, alkenyl, substitutedalkenyl, halo, alkoxyl, heteroalkyl, substituted heteroalkyl,heteroalkenyl, or substituted heteroalkenyl , or a salt thereof. In someembodiments, the X is hydrogen (i.e., the dihydromyricetin compound isdihydromyricetin). In some embodiments, the X is independently selectedfrom the group consisting of —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —CH(CH₃)₂,—CH₂CH₂CH₂CH₃, —CH₂CH(CH₃)₂, —C(CH₃)₃, —CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₃,—C(CH₃)₂CH₂CH₃, —CH₂CH₂CH(CH₃)₂, —CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃,—CH₂CH₂C(CH₃)₂CH₃, —CH₂CH₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃,—CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₂CH₃, and—CH₂CH₂CH₂CH₂OCH₂CH₃. In some embodiments, the dihydromyricetin compoundis of formula (II):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is of formula (III):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments of the methods described above, the dihydromyricetincompound is provided in a hair product (e.g., a shampoo, a conditioner,or a hair spray, etc.). In some embodiments, the dihydromyricetincompound in the hair product is in a concentration of about 0.01 μM toabout 250 μM. In some embodiments, the dihydromyricetin compound in thehair product is in a concentration of about 0.5 μM to about 10 μM. Insome embodiments, the dihydromyricetin compound is synthetic orextracted and purified from a whole plant or a plant tissue of Hoveniadulcis, Leptarrhena pyrolifolia, Pinus contorta, Ampelopsisgrossedentata, Glochidion sumatranum, Rhododendron ferrugineum, Ericaarborea, Salix hulteni, Manilkara zapota, Catharanthus roseus, Myricarubra, or Xanthoceras sorbifolia.

In some embodiments of the methods described above, the individual is ahuman. In some embodiments, the individual has chronic stress, alopecia,and/or baldness.

In another aspect, the invention provides a method for delaying orreversing signs of aging skin comprising applying an effective amount ofa dihydromyricetin compound to the skin of an individual, wherein thedihydromyricetin compound is of formula (I):

wherein each X is independently selected from hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, halo, alkoxyl,heteroalkyl, substituted heteroalkyl, heteroalkenyl, or substitutedheteroalkenyl, or a salt thereof. In some embodiments, the X is hydrogen(i.e., the dihydromyricetin compound is dihydromyricetin). In someembodiments, the X is independently selected from the group consistingof —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —CH(CH₃)₂, —CH₂CH₂CH₂CH₃, —CH₂CH(CH₃)₂,—C(CH₃)₃, —CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₃, —C(CH₃)₂CH₂CH₃,—CH₂CH₂CH(CH₃)₂, —CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃,—CH₂CH₂C(CH₃)₂CH₃, —CH₂CH₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃,—CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₂CH₃, and—CH₂CH₂CH₂CH₂OCH₂CH₃. In some embodiments, the dihydromyricetin compoundis of formula (II):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is of formula (III):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is extracted andpurified from a whole plant or a plant tissue of Hovenia dulcis,Leptarrhena pyrolifolia, Pinus contorta, Ampelopsis grossedentata,Glochidion sumatranum, Rhododendron ferrugineum, Erica arborea, Salixhulteni, Manilkara zapota, Catharanthus roseus, or Xanthocerassorbifolia.

In some embodiments, the dihydromyricetin compound is provided in a skinproduct (e.g., a moisturizer, a facial polisher, a facial cleaner, asunscreen, or a skin patch, etc.). In some embodiments, thedihydromyricetin compound in the skin product is in a concentration ofabout 0.01 μM to about 250 μM. In some embodiments, the dihydromyricetincompound in the skin product is in a concentration of about 0.5 μM toabout 10 μM. In some embodiments, the dihydromyricetin compound in theskin product is in a concentration of at least about 5% (w/v-%) or atleast about 5% (w/w-%). In some embodiments, the dihydromyricetincompound in the skin product is in a concentration of at least about 10%(w/v-%) or at least about 10% (w/w-%).

In some embodiments, the individual is a human. In some embodiments, theindividual has chronic stress. In some embodiments, the individual hasskin rash, skin dehydration, or psycho-emotional changes associated withchronic stress.

In another aspect, the invention provides a cosmetic product comprisinga dihydromyricetin compound or a salt thereof, wherein thedihydromyricetin compound is of formula (I):

wherein each X is independently selected from hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, halo, alkoxyl,heteroalkyl, substituted heteroalkyl, heteroalkenyl, or substitutedheteroalkenyl, and wherein said dihydromyricetin in the product is atleast about 0.01 μM. In some embodiments, the X is hydrogen (i.e., thedihydromyricetin compound is dihydromyricetin). In some embodiments, theX is independently selected from the group consisting of —CH₃, —CH₂CH₃,—CH₂CH₂CH₃, —CH(CH₃)₂, —CH₂CH₂CH₂CH₃, —CH₂CH(CH₃)₂, —C(CH₃)₃,—CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₃, —C(CH₃)₂CH₂CH₃, —CH₂CH₂CH(CH₃)₂,—CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃, —CH₂CH₂C(CH₃)₂CH₃,—CH₂CH₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃, —CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃,—CH₂CH₂CH₂OCH₂CH₃, and —CH₂CH₂CH₂CH₂OCH₂CH₃. In some embodiments, thedihydromyricetin compound is of formula (II):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is of formula (III):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound in the product isabout 0.01 μM to about 250 μM. In some embodiments, the dihydromyricetincompound in the product is about 0.5 μM to about 10 μM.

In another aspect, the invention provides a cosmetic product comprisinga dihydromyricetin compound or a salt thereof, wherein thedihydromyricetin compound is of formula (I):

wherein each X is independently selected from hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, halo, alkoxyl,heteroalkyl, substituted heteroalkyl, heteroalkenyl, or substitutedheteroalkenyl, and wherein purified dihydromyricetin has been added tothe cosmetic product or a material from which the cosmetic product ismade. In some embodiments, the X is hydrogen (i.e., the dihydromyricetincompound is dihydromyricetin). In some embodiments, the X isindependently selected from the group consisting of —CH₃, —CH₂CH₃,—CH₂CH₂CH₃, —CH(CH₃)₂, —CH₂CH₂CH₂CH₃, —CH₂CH(CH₃)₂, —C(CH₃)₃,—CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₃, —C(CH₃)₂CH₂CH₃, —CH₂CH₂CH(CH₃)₂,—CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃, —CH₂CH₂C(CH₃)₂CH₃,—CH₂CH₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃, —CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃,—CH₂CH₂CH₂OCH₂CH₃, and —CH₂CH₂CH₂CH₂OCH₂CH₃. In some embodiments, thedihydromyricetin compound is of formula (II):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is of formula (III):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments of the cosmetic product described above, thedihydromyricetin compound is synthetic or extracted and purified from awhole plant or a plant tissue of Hovenia dulcis, Leptarrhenapyrolifolia, Pinus contorta, Ampelopsis grossedentata, Glochidionsumatranum, Rhododendron ferrugineum, Erica arborea, Salix hulteni,Manilkara zapota, Catharanthus roseus, Myrica rubra, or Xanthocerassorbifolia and added to the cosmetic product or the material from whichthe cosmetic product is made. In some embodiments of the cosmeticproduct described above, the purified dihydromyricetin compound is atleast about 90% pure, at least about 95% pure, at least about 98% pure,or at least about 99% pure.

In some embodiments of the cosmetic product described above, thecosmetic product is a hair product (e.g., a shampoo, a conditioner, or ahair spray, etc.). In some embodiments of the cosmetic product describedabove, the cosmetic product is a skin product (e.g., a moisturizer, afacial polisher, a facial cleaner, a sunscreen, or a skin patch, etc.).

In another aspect, the invention provides a method of making a cosmeticproduct comprising adding a dihydromyricetin compound or a salt thereofto a cosmetic product or a material from which the cosmetic product ismade, wherein the dihydromyricetin compound is of formula (I):

wherein each X is independently selected from hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, halo, alkoxyl,heteroalkyl, substituted heteroalkyl, heteroalkenyl, or substitutedheteroalkenyl, and wherein the final concentration of thedihydromyricetin compound in the cosmetic product is at least about 0.01μM. In some embodiments, the X is hydrogen (i.e., the dihydromyricetincompound is dihydromyricetin). In some embodiments, the X isindependently selected from the group consisting of —CH₃, —CH₂CH₃,—CH₂CH₂CH₃, —CH(CH₃)₂, —CH₂CH₂CH₂CH₃, —CH₂CH(CH₃)₂, —C(CH₃)₃,—CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₃, —(CH₃)₂CH₂CH₃, —CH₂CH₂CH(CH₃)₂,—CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃, —CH₂CH₂C(CH₃)₂CH₃,—CH₂Ch₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃, —CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃,—CH₂CH₂CH₂OCH₂CH₃, and —CH₂CH₂CH₂CH₂OCH₂CH₃. In some embodiments, thedihydromyricetin compound is of formula (II):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is of formula (III):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the final concentration of the dihydromyricetincompound in the cosmetic product is about 0.01 μM to about 250 μM. Insome embodiments, the final concentration of the dihydromyricetincompound in the cosmetic product is about 0.5 μM to about 10 μM.

In another aspect, the invention provides a method of making a cosmeticproduct comprising adding a purified dihydromyricetin compound or a saltthereof to a cosmetic product or a material from which the cosmeticproduct is made, wherein the dihydromyricetin compound is of formula(I):

wherein each X is independently selected from hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, halo, alkoxyl,heteroalkyl, substituted heteroalkyl, heteroalkenyl, or substitutedheteroalkenyl, and wherein the final concentration of thedihydromyricetin compound in the cosmetic product is at least about 0.01μM. In some embodiments, the X is hydrogen (i.e., the dihydromyricetincompound is dihydromyricetin). In some embodiments, the X isindependently selected from the group consisting of —CH₃, —CH₂CH₃,—CH₂CH₂CH₃, —CH(CH₃)₂, —CH₂CH₂CH₂CH₃, —CH₂CH(CH₃)₂, —C(CH₃)₃,—CH₂CH₂CH₂CH₂CH₃, ——CH₂C(CH₃)₂CH₃, —C(CH₃)₂CH₂CH₃, —CH₂CH₂CH(CH₃)₂,—CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃, —CH₂CH₂C(CH₃)₂CH₃,—CH₂CH₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃, —CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃,—CH₂CH₂CH₂OCH₂CH₃, and —CH₂CH₂CH₂CH₂OCH₂CH₃. In some embodiments, thedihydromyricetin compound is of formula (II):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is of formula (III):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments of the method of making a cosmetic product describedabove, the dihydromyricetin compound is synthetic or extracted andpurified from a whole plant or a plant tissue of Hovenia dulcis,Leptarrhena pyrolifolia, Pinus contorta, Ampelopsis grossedentata,Glochidion sumatranum, Rhododendron ferrugineum, Erica arborea, Salixhulteni, Manilkara zapota, Catharanthus roseus, Myrica rubra, orXanthoceras sorbifolia. In some embodiments, the purifieddihydromyricetin compound is at least about 90% pure, at least about 95%pure, at least about 98% pure, or at least about 99% pure.

In some embodiments of the method of making a cosmetic product describedabove, the cosmetic product is a hair product (e.g., a shampoo, aconditioner, or a hair spray, etc.). In some embodiments of the methodof making a cosmetic product described above, the cosmetic product is askin product (e.g., a moisturizer, a facial polisher, a facial cleaner,a sunscreen, or a skin patch, etc.).

In another aspect, the invention provides a method of extracting andpurifying dihydromyricetin from a whole plant or a plant tissuecomprising the steps of (a) extracting a whole plant or a plant tissuethat contains dihydromyricetin with a first ethanol water solution; (b)concentrating and crystalizing compounds in the extract of step (a); (c)re-extracting the crystalized compounds with a second ethanol watersolution; and (d) concentrating and crystalizing compounds in theextract of step (c), wherein the crystalized material in step (d)contains purified dihydromyricetin. In some embodiments, the firstethanol water solution contains about 60% to about 80% ethanol (such asabout 70% ethanol). In some embodiments, the second ethanol watersolution contains about 70% to about 90% ethanol (such as about 80%ethanol). In some embodiments, the extraction is from a whole plant or aplant tissue of Hovenia dulcis, Leptarrhena pyrolifolia, Pinus contorta,Ampelopsis grossedentata, Glochidion sumatranum, Rhododendronferrugineum, Erica arborea, Salix hulteni, Manilkara zapota,Catharanthus roseus, Myrica rubra, or Xanthoceras sorbifolia. In someembodiments, the crystalized material in step (d) contains at least 95%or at least 98% dihydromyricetin.

In another aspect, the invention provides kits or articles ofmanufacture comprising (a) a composition comprising a dihydromyricetincompound, wherein the dihydromyricetin compound is of formula (I):

wherein each X is independently selected from hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, halo, alkoxyl,heteroalkyl, substituted heteroalkyl, heteroalkenyl, or substitutedheteroalkenyl, or a salt thereof and (b) a package insert or a labelindicating that the product is useful for promoting hair growth,reversing hair loss (or balding), promoting natural hair regrowth,increasing the thickness of thin (or thinning) hair, or preventing hairloss (or balding). In some embodiments, the X is hydrogen (i.e., thedihydromyricetin compound is dihydromyricetin). In some embodiments, theX is independently selected from the group consisting of —CH₃, —CH₂CH₃,—CH₂CH₂CH₃, —CH(CH₃)₂, —CH₂CH₂CH₂CH₃, —CH₂CH(CH₃)₂, —C(CH₃)₃,—CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₃, —C(CH₃)₂CH₂CH₃, —CH₂CH₂CH(CH₃)₂,—CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃, —CH₂CH₂C(CH₃)₂CH₃,—CH₂CH₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃, —CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃,—CH₂CH₂CH₂OCH₂CH₃, and —CH₂CH₂CH₂CH₂OCH₂CH₃. In some embodiments, thedihydromyricetin compound is of formula (II):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is of formula (III):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In another aspect, the invention provides kits or articles ofmanufacture comprising (a) a composition comprising a dihydromyricetincompound, wherein the dihydromyricetin compound is of formula (I):

wherein each X is independently selected from hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, halo, alkoxyl,heteroalkyl, substituted heteroalkyl, heteroalkenyl, or substitutedheteroalkenyl, or a salt thereof; and (b) a package insert or a labelindicating that the product is useful for delaying skin aging orreversing the signs of skin aging. In some embodiments, the X ishydrogen (i.e., the dihydromyricetin compound is dihydromyricetin). Insome embodiments, the X is independently selected from the groupconsisting of —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —CH(CH₃)₂, —CH₂CH₂CH₂CH₃,—CH₂CH(CH₃)₂, —C(CH₃)₃, —CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₃,—C(CH₃)₂CH₂CH₃, —CH₂CH₂CH(CH₃)₂, —CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃,—CH₂CH₂C(CH₃)₂CH₃, —CH₂CH₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃,—CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₂CH₃, and—CH₂CH₂CH₂CH₂OCH₂CH₃. In some embodiments, the dihydromyricetin compoundis of formula (II):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the dihydromyricetin compound is of formula (III):

wherein each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.

In some embodiments, the composition comprising the dihydromyricetincompound is a cosmetic product. In some embodiments, the compositioncomprising the dihydromyricetin compound is packaged in a container withthe insert or a label. In some embodiments, the label further indicatesthat the product has a stress reducing, calming, and/or soothing effect.

It is to be understood that one, some, or all of the properties of thevarious embodiments described herein may be combined to form otherembodiments of the present invention. These and other aspects of theinvention will become apparent to one of skill in the art.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows that a chronically stressed mouse exhibited hair loss.

FIG. 2 shows that chronically stressed mice exhibited behavioralanxiety. FIG. 2A shows the results of mice that were observed in theopen arms in an elevated plus maze (EPM-open) behavioral test. FIG. 2Bshows the results of mice that were observed in the enclosed arms in anelevated plus maze (EPM-closed). FIG. 2C shows the results of the amountof time mice avoided the open center in an open field test (OFT)behavioral test. FIG. 2D shows the results of locomotor activity in anOFT behavioral test.

FIG. 3 shows the regrowth of hair on chronically stressed mice thatunderwent daily treatment with 1 μM dihydromyricetin for a period of 6weeks.

FIG. 4 shows that behavioral anxiety was reduced in chronically stressedmice that underwent daily treatment with 1 μM dihydromyricetin for aperiod of 6 weeks. FIG. 4A shows the results of mice that were observedin the open arms in an elevated plus maze (EPM-open) behavioral test.FIG. 4B shows the results of mice that were observed in the enclosedarms in an elevated plus maze (EPM-closed). FIG. 4C shows the results ofthe amount of time mice avoided the open center in an open field test(OFT) behavioral test. FIG. 4D shows the results of locomotor activityin an OFT behavioral test.

FIG. 5 shows the effects of vehicle (negative control), DHM 5%, DHM 10%,and positive control (Minoxidil 2%) on the induction of anagen on theskin of mice after 8 days of topical application. Statisticalsignificance was evaluated with the non-parametric Kruskal-Walis testfollowed by Dunn's multiple comparison test. * P<0.05. N=10 per group.

FIG. 6 shows the effects of vehicle (negative control), DHM 5%, DHM 10%,and positive control (Minoxidil 2%) on hair growth of mice after 12 daysof topical application. Statistical significance was evaluated with thenon-parametric Kruskal-Walis test followed by Dunn's multiple comparisontest. *P<0.05. N=10 per group.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides, inter alia, methods of treating hair loss,methods of inducing hair growth, method of preventing hair loss, methodsof delaying signs of aging skin, and methods of treating chronic stressby administering an effective amount of dihydromyricetin, or aderivative thereof. The invention also provides cosmetic productscomprising at least 0.5 μM dihydromyricetin and/or purifieddihydromyricetin, or a derivative thereof; and methods of making suchcosmetic products.

The methods and compositions of the invention are based on theunexpected observation that applying dihydromyricetin, a naturalflavonol extracted from plants, to the sites of hair loss induces newhair growth. Moreover, dihydromyricetin use does not produce theunpleasant side effects associated with other topical hair growthtreatments, e.g., dryness, irritation, and allergic reaction.

A. Definitions

As used herein, a “dihydromyricetin compound” refers todihydromyricetin, a dihydromyricetin derivative, or a salt thereof.

As used herein, “treating or preventing hair loss” refers to the abilityto reduce, reverse, slow, or prevent symptoms of hair loss, including,but not limited to, alopecia, hair loss, thinning hair, uneven thicknessof hair, and/or bald patches associated with, e.g., adrenergic alopecia,telogen effluvium, alopecia areata, traumatic alopecia, anageneffluvium, and hair loss associated with nutritional deficiencies,metabolic defects, marked weight loss, and chronic stress. Treating orpreventing hair loss also includes promoting growth at the site(s) ofthe hair loss.

As used herein, to “induce hair growth” means to promote hair growth orre-growth or increase the amount of hair (such as new hair growth). Forexample, inducing hair growth encompasses promotion or stimulation ofhair growth in general (e.g., to thicken naturally thin or thinninghair) or at the site(s) of baldness or hair loss.

As used herein, to “delaying or reversing signs of aging skin” means todefer, hinder, slow, retard, stabilize, and/or postpone development ofsymptoms of skin aging, including, but not limited to, the appearance ofwrinkles, sagging skin, thinning skin, cornification, elastosis, theappearance of hyperpigmented spots (i.e., freckles, age spots, or “liverspots”), and/or dryness. This delay can be of varying lengths of time,depending on the condition of the skin and/or individual being treated.

As used herein, an “effective amount” refers to at least an amounteffective, at dosages and for periods of time necessary, to achieve thedesired result, e.g., induction of hair growth, prevention of hair loss,delaying or reversing signs of aging skin, or reduction in stress. Aneffective amount can be provided in one or more administrations.

As used herein, “alkyl” refers to a univalent group derived from asaturated hydrocarbon by removing one hydrogen atom. The saturatedhydrocarbon may contain normal, secondary, or tertiary carbon atoms.These carbon atoms may be arranged in straight or branched chain, or incyclic ring, or a combination thereof. For example, an alkyl group canhave 1 to 20 carbon atoms (i.e., C1-C20 alkyl). 1 to 12 carbon atoms(i.e., C1-C12 alkyl), or 1 to 6 carbon atoms (i.e., C1-C6 alkyl).Examples of suitable alkyl groups include, but are not limited to,methyl (Me, —CH3), ethyl (Et, CH2CH3). 1-propyl (n-Pr, n-propyl,—CH2CH2CH3), 2-propyl Q-Pr, i-propyl, —CH(CHs)2), 1-butyl (n-Bu.n-butyl, —CH2CH2CH2CH3), 2-methyl-1-propy I Q-Bu, i-butyl,—ClhCH(CH₃)₂), 2butyl (s-Bu, s-butyl, —CH(CH₃)CH₂CH₃). 2-methyl-2-propyl(t-Bu, t-butyl, —C(CH_(s))₃), 1-penty (n-pentyl, —CH₂CH₂CH₂CH₂CH₃),2-pentyl (—CH(CH₃)CH₂CH₂CH₃), 3-pentyl (—CH(CH₂CH₃)₂). 2-methyl-2-butyl(—C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl (—CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl(—CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl (—CH₂CH(CH₃)CH₂CH₃). 1-hexyl(CH₂CH₂CH₂CH₂CH₂CH₃), 2-hexyl (—CH(CH₃)CH₂CH₂CH₂CH₃), 3-hexyl(CH(CH₂CH₃)(CH₂CH₂CH₃)), 2-methyl-2-pentyl (-C(CH₃)₂CH₂CH₂CH₃),3-methyl-2-pentyl (CH(CH₃)CH(CH₃)CH₂CH₃), 4-methyl-2-pentyl(—CH(CH₃)CH₂CH(CH₃)₂), 3-methyl-3-pentyl (C(CH₃)(CH₂CH₃)₂),2-methyl-3-pentyl (—CH(CH₂CH₃)CH(CH₃)₂), 2,3-dimethyl-2-butyl(C(CH₃)₂CH(CH₃)₂), 3,3-dimethyl-2-butyl (—CH(CH₃)C(CH₃)₃, and octyl(—(CH₂)₇CH₃).

As used herein, “alkylene” refers to a divalent group derived from analkyl by removing one hydrogen atom. That is, “alkylene” can be asaturated, branched or straight chain or cyclic hydrocarbon radicalhaving two monovalent radical centers derived by the removal of twohydrogen atoms from the same or two different carbon atoms of a parentalkane. For example, an alkylene group can have 1 to 20 carbon atoms, 1to 12 carbon atoms, or 1 to 6 carbon atoms. Typical alkylene radicalsinclude, but are not limited to, methylene (—CH₂—), 1,1-ethyl(—CH(CH₃)—), 1,2-ethyl (—CH₂CH₂—), 1,1-propyl (—CH(CH₂CH₃)—), 1,2-propyl(—CH₂CH(CH₃)—), 1,3-propyl (—CH₂CH₂CH₂—), 1,4butyl (—CH₂CH₂CH₂CH₂—), andthe like.

As used herein, “alkenyl” refers to a univalent group derived from ahydrocarbon by removing one hydrogen atom wherein the hydrocarboncontains at least one carbon-to-carbon double bond. For example, analkenyl group can have 1 to 20 carbon atoms (i.e., C1-C20 alkenyl), 1 to12 carbon atoms (i.e., C1-C12 alkenyl), or 1 to 6 carbon atoms (i.e.,C1-C6 alkenyl). Typical alkenyl groups include, but are not limited to,ethenyl, prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl (allyl),cycloprop-1-en-1-yl; cycloprop-2-en-1-yl, but-1-en-1 yl, but-1-en-2-yl,2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-2-yl,buta-1,3--dien-1-yl, butal, 3-dien-2-yl, cyclobut-1-en-1-yl,cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl, and the like.

As used herein, “alkenylene” refers to a divalent group derived from analkenyl by removing one hydrogen atom. That is, “alkenylene” can be anunsaturated, branched or straight chain or cyclic unsaturatedhydrocarbon radical having two monovalent radical centers derived by theremoval of two hydrogen atoms from the same or two different carbonatoms of a parent alkene.

As used herein, “alkoxyl” refers to a monovalent radical -OR wherein Ris an alkyl or alkenyl.

As used herein, “halo” refers to a univalent group derived from ahalogen element including, but not limited to, fluorine, chlorine,bromine, iodine, and astatine.

As used herein, “heteroalkyl” or “heteroalkenyl” refers to alkyl oralkenyl, respectively, in which one or more of the carbon atoms (andoptionally any associated hydrogen atoms), are each, independently ofone another, replaced with the same or different heteroatoms orheteroatomic groups. Similarly, “heteroalkylene” or “heteroalkenylene”refers to alkylene or alkenylene, respectively, in which one or more ofthe carbon atoms (and optionally any associated hydrogen atoms), areeach, independently of one another, replaced with the same or differentheteroatoms or heteroatomic groups. Typical heteroatoms or heteroatomicgroups which can replace the carbon atoms include, but are not limitedto, —O—, —S—, —N—, —Si—, —NH—, —S(O)—, —S(O)₂—, —S(O)NH—, —S(O)₂NH— andthe like, and combinations thereof. The heteroatoms or heteroatomicgroups may be placed at any interior position of the alkyl or alkenyl.Typical heteroatomic groups which can be included in these groupsinclude, but are not limited to, —O—, —S—, —O—O, —S—S—, —O—S—,—N(R^(a))₂—, ═N—N═, —N═N—, —N═N—N(R^(a))₂, —PR^(a)—, —P(O)₂—, —POR^(a)—,—O—P(O)₂—, —SO—, —SO₂—, —Sn(R^(a))₂—0 and the like, where each R^(a) isindependently hydrogen, alkyl, substituted alkyl, aryl, substitutedaryl, arylalkyl, substituted arylalkyl, cycloalkyl, substitutedcycloalkyl, cyclohetero alkyl, substituted cycloheteroalkyl,heteroalkyl, substituted heteroalkyl, heteroaryl, substitutedheteroaryl, heteroarylalkyl or substituted heteroarylalkyl, or aprotecting group.

As used herein, “protecting group” refers to a grouping of atoms thatwhen attached to a reactive functional group in a molecule masks,reduces or prevents reactivity of the functional group. Representativeamino protecting groups include, but are not limited to, formyl, acetyl,trifiuoroacetyl, benzyl, benzyloxycarbonyl (“CBZ”), tertbutoxycarbonyl(“Boc”), trimethylsilyl (“TMS”), 2-trimethylsilyl-ethanesulfonyl(“SES”), trityl and substituted trityl groups, allyloxycarbonyl,9-fiuorenylmethyloxycarlbonyl (“FMOC”), nitroveratryloxycarbonyl(“NVOC”), and the like. Representative hydroxy protecting groupsinclude, but are not limited to, those where the hydroxy group is eitheracylated or alkylated such as benzyl, and trityl ethers as well as alkylethers, tetrahydropyranyl ethers, trialkylsilyl ethers and allyl ethers.

As used here, “substituted,” when used to modify a specified group orradical, refers to one or more hydrogen atoms of the specified group orradical are each, independently of one another, replaced with the sameor different substituent(s). Substituent groups useful for substitutingsaturated carbon atoms in the specified group or radical include, butare not limited to, —R⁰, halo, O^(a), ═O, —OR^(b), —SR^(b), —S^(a), ═S,—N(R^(d))₂, ═NR^(b), ═N—OR^(b) tnhalomethyl, —CF₃, —CN, —OCN, —SCN, NO,—NO₂, ═N₂, —N₃, —S(O)₂R^(b), —S(O)₂NR^(b), —S(O)₂O^(a), —S(O)₂OR^(b),—OS(O)₂R^(b), —OS(O)₂O^(n), —OS(O)₂OR^(b), —P(O)(O)₂,—P(O)(OR^(b))(O^(n)), —P(O)(OR^(b))(OR^(b)), —C(O)R^(b), —C(S)R^(b),—C(NR^(b))R^(b), C(O)O^(n), —C(O)OR^(b), —C(S)OR^(b), —C(O)N(R^(d))₂),—C(NR^(b))N(R(^(d))_(2′), —OC(O)R^(b), —OC(S)R^(b), —OC(O)O^(n),OC(O)OR^(b), —OC(S)OR^(b), —NR^(b)C(O)R^(b), —NR^(b)C(S)R^(b),—NR^(b)C(O)O^(a), —NR^(b)C(O)OR^(b), —NR^(b)C(S)OR^(b).—NR⁶C(O)N(R^(d))₂, —NR⁶C(NR⁶)R^(b), and —NR⁶C(NR⁶)N(R^(d))₂, where R^(c)is selected from alkyl, cycloalkyl, heteroalkyl, cycloheteroalkyl, aryl,arylalkyl, heteroaryl and heteroarylalkyl; each R^(b) is independentlyhydrogen, a protecting group, or R^(o); and each R^(d) is independentlyR^(b) or alternatively, the two R^(d)s may be taken together with thenitrogen atom to which they are bonded form a 4-, 5-, 6-or 7-memberedcycloheteroalkyl which may optionally include from 1 to 4 of the same ordifferent additional heteroatoms selected from O, N, and S. As specificexamples, —N(R^(d))₂ is meant to include —NH₂, —NH-alkyl,N-pyrrolidinyl, and N-morpholinyl. As another specific example, asubstituted alkyl is meant to include -alkylene-O-alkyl,-alkylene-heteroaryl, -alkylene-cycloheteroalkyl, -alkyleneC(O)OR^(b),-alkylene-C(O)N(R^(d))₂, and —CH₂—CH₂—C(O)—CH₃. The one or moresubstituent groups, taken together with the atoms to which they arebonded, may form a cyclic ring including cycloalkyl andcycloheteroalkyl.

As used herein, the term “w/v-%” refers to the weight of a compound,such as dihydromyricetin, (in grams) for every 100 ml of a liquidproduct of the present disclosure containing such a compound, such as ahair product or other cosmetic products.

As used herein, the term “w/w-%” refers to the weight of a compound,such as dihydromyricetin, (in grams) for every gram of a product of thepresent disclosure containing such compound, such as a hair product orother cosmetic products.

An “individual” or a “subject” is a mammal, more preferably a human.Mammals also include, but are not limited to, farm animals, sportanimals, pets (such as cats, dogs, horses), primates, mice and rats.

As used herein, the singular form “a”, “an”, and “the” includes pluralreferences unless indicated otherwise.

Reference to “about” a value or parameter herein refers to the usualerror range for the respective value readily known to the skilled personin this technical field. Reference to “about” a value or parameterherein includes (and describes) aspects that are directed to that valueor parameter per se. For example, description referring to “about X”includes description of “X.”

It is understood that aspects and embodiments of the invention describedherein include “comprising,” “consisting,” and “consisting essentiallyof” aspects and embodiments.

B. Methods of Treating or Preventing Hair Loss, Promoting Hair Growth,Delaying the Signs of Aging Skin or Reversing the Signs of Aging Skin

In one aspect, the invention provides methods for treating hair loss orinducing hair growth, and methods of preventing hair loss, that includeapplying an effective amount of a dihydromyricetin compound, such asdihydromyricetin, to the scalp of an individual. In certain embodimentsof the methods, the dihydromyricetin compound is applied to the scalp ofthe individual through any known method, including but not limited totopically spreading, spraying, steaming, soaking, washing, etc. Themethods can also be useful for other sites of hair loss or sites inwhich hair growth is desired, such as the chin (to promote, even out, orthicken beard growth), the sides of the face (to promote, even out, orthicken the growth of sideburns), between the nose and lip (to promote,even out, or thicken moustache growth), the chest, etc. An effectiveamount of the dihydromyricetin compound may be applied to these sites.

Another aspect of the invention provides methods for treating hair lossor inducing hair growth, and methods of preventing hair loss, thatinclude applying an effective amount of a dihydromyricetin compound,such as dihydromyricetin, to the skin of an animal. In certainembodiments of the methods, the dihydromyricetin compound is applied tothe skin of the animal through any known method, including but notlimited to topically spreading, spraying, steaming, soaking, washing,etc. In certain embodiments, the animal is a pet animal, includingwithout limitation, a dog, a cat, a horse, a rabbit, a hamster, a guineapig, a hamster, etc.

In certain embodiments of the methods, the dihydromyricetin compound isprovided in a hair product. In certain embodiments of the methods, thehair products that comprise the dihydromyricetin compound include, butare not limited to, e.g., shampoos, conditioners, masks, sprays ormists, gels, mousses, foams, serums, pastes, pomades, powders, oils,emulsions, creams, waxes, glazes, balms, tonics, lotions, ointments,polishes, lightening agents, straightening agents, relaxing agents,curling agents, or dyes. In certain embodiments of the methods, thedihydromyricetin compound in the hair product is at a concentration ofabout 0.01 μM to about 250 μM. For example, the dihydromyricetincompound in the hair product is at a concentration of about 0.01 μM,about 0.05 μM, about 0.1 μM, about 0.2 μM, about 0.3 μM, about 0.4 μM,0.5 μM, about 0.6 μM, about 0.7 μM, about 0.8 μM, about 0.9 μM, about 1μM, about 2 μM, about 3 μM, about 4 μM, about 5 μM, about 6 μM, about 7μM, about 8 μM, about 9 μM, about 10 μM, about 25 μM, about 50 μM, about100 μM, about 150 μM, about 200 μM, or about 250 μM, including any rangein between these values.

In certain embodiments of the methods, the dihydromyricetin compound inthe hair product is at a concentration of at least about 5% (w/v-%) orat least about 5% (w/w-%) to at least about 25% (w/v-%) or at leastabout 25% (w/w-%). For example, the dihydromyricetin compound in thehair product is at a concentration of about at least about 5% (w/v-%) orat least about 5% (w/w-%), at least about 6% (w/v-%) or at least about6% (w/w-%), at least about 7% (w/v-%) or at least about 7% (w/w-%), atleast about 8% (w/v-%) or at least about 8% (w/w-%), at least about 9%(w/v-%) or at least about 9% (w/w-%), at least about 10% (w/v-%) or atleast about 10% (w/w-%), at least about 11% (w/v-%) or at least about11% (w/w-%), at least about 12% (w/v-%) or at least about 12% (w/w-%),at least about 13% (w/v-%) or at least about 13% (w/w-%), at least about14% (w/v-%) or at least about 14% (w/w-%), at least about 15% (w/v-%) orat least about 15% (w/w-%), at least about 16% (w/v-%) or at least about16% (w/w-%), at least about 17% (w/v-%) or at least about 17% (w/w-%),at least about 18% (w/v-%) or at least about 18% (w/w-%), at least about19% (w/v-%) or at least about 19% (w/w-%), at least about 20% (w/v-%) orat least about 20% (w/w-%), at least about 21% (w/v-%) or at least about21% (w/w-%), at least about 22% (w/v-%) or at least about 22% (w/w-%),at least about 23% (w/v-%) or at least about 23% (w/w-%), at least about24% (w/v-%) or at least about 24% (w/w-%), or at least about 25% (w/v-%)or at least about 25% (w/w-%), including any range in between thesevalues.

In certain embodiments, the compound in a relatively higherconcentration is applied to treat hair loss or induce hair growth, andthe compound in a relatively lower concentration is applied to maintainthe amount of hair. In some embodiments, “preventing hair loss” includesmaintaining hair growth and/or the amount of hair resulting from hairgrowth (such as induced hair growth). Also provided herein is a methodof maintaining the amount of hair comprising applying an effectiveamount of a dihydromyricetin compound described herein to scalp of anindividual or other sites.

In certain embodiments of the methods, the individual suffers fromchronic stress, alopecia, thinning hair, bald patches, and/or baldness.In certain embodiments, alopecia, thinning hair, hair loss, baldpatches, and/or baldness is associated with chronic stress and/oranxiety. In certain embodiments of the methods, the individual hastelogen effluvium (TE) or alopecia areata.

In another aspect, the invention provides methods for delaying orreversing signs of aging skin comprising applying an effective amount ofa dihydromyricetin compound, such as dihydromyricetin, to the skin of anindividual, including, but not limited to, e.g., to the face, the neck,the decolletage, the hands, the arms, the shoulders, the legs, etc. Incertain embodiments, the dihydromyricetin compound delays or reversesthe appearance of wrinkles and folds, scaliness, flakiness, roughnessand/or other forms of uneven skin texture, sagging skin, thinning skin,cornification, elastosis or loss of skin elasticity, discoloration (suchas under eye circles), blotching, sallowness, hyperpigmented skinregions (such as age spots, “liver spots”, and/or freckles), keratoses,solar purpuras, hyperkeratinization, telangiectasia (i.e., “spidervessels”), and/or dryness.

In certain embodiments of the methods, the dihydromyricetin compound isprovided in a skin product. In certain embodiments of the methods, theskin product containing the dihydromyricetin compound is a lotion, amoisturizer, a facial polisher, a facial cleanser, a sunscreen, a skinpatch, a scrub or exfoliating product, an astringent, a toner, a mask, apeel, a gel, a cream, a balm, a wax, an oil, a salve, a makeup remover,an insect repellent, a soap, a makeup product (e.g., a foundation, aprimer, a concealer or color corrector, a blusher or rouge, a lipstick,a lip gloss, a lip balm, a bronzer, a powder, a setting spray, etc.), amist or spray, an ointment, a liniment, a topical analgesic, a topicalantihistamine, or an emulsion. In certain embodiments of the methods,the dihydromyricetin compound in the skin product is at a concentrationof about 0.01 μM to about 250 μM. For example, the dihydromyricetincompound in the skin product is at a concentration of about 0.01 μM,about 0.05 μM, about 0.1 μM, about 0.2 μM, about 0.3 μM, about 0.4 μM,0.5 μM, about 0.6 μM, about 0.7 μM, about 0.8 μM, about 0.9 μM, about 1μM, about 2 μM, about 3 μM, about 4 μM, about 5 μM, about 6 μM, about 7μM, about 8 μM, about 9 μM, about 10 μM, about 25 μM, about 50 μM, about100 μM, about 150 μM, about 200 μM, or about 250 μM, including any rangein between these values. In certain embodiments of the methods, thedihydromyricetin compound in the skin product is at a concentration ofat least about 5% (w/v-%) or at least about 5% (w/w-%) to at least about25% (w/v-%) or at least about 25% (w/w-%). For example, thedihydromyricetin compound in the skin product is at a concentration ofabout at least about 5% (w/v-%) or at least about 5% (w/w-%), at leastabout 6% (w/v-%) or at least about 6% (w/w-%), at least about 7% (w/v-%)or at least about 7% (w/w-%), at least about 8% (w/v-%) or at leastabout 8% (w/w-%), at least about 9% (w/v-%) or at least about 9%(w/w-%), at least about 10% (w/v-%) or at least about 10% (w/w-%), atleast about 11% (w/v-%) or at least about 11% (w/w-%), at least about12% (w/v-%) or at least about 12% (w/w-%), at least about 13% (w/v-%) orat least about 13% (w/w-%), at least about 14% (w/v-%) or at least about14% (w/w-%), at least about 15% (w/v-%) or at least about 15% (w/w-%),at least about 16% (w/v-%) or at least about 16% (w/w-%), at least about17% (w/v-%) or at least about 17% (w/w-%), at least about 18% (w/v-%) orat least about 18% (w/w-%), at least about 19% (w/v-%) or at least about19% (w/w-%), at least about 20% (w/v-%) or at least about 20% (w/w-%),at least about 21% (w/v-%) or at least about 21% (w/w-%), at least about22% (w/v-%) or at least about 22% (w/w-%), at least about 23% (w/v-%) orat least about 23% (w/w-%), at least about 24% (w/v-%) or at least about24% or at least about 25% (w/v-%) or at least about 25% (w/w-%),including any range in between these values.

In certain embodiments of the methods, the individual suffers fromchronic stress. In certain embodiments of the methods, the individualhas a skin rash, has dehydrated skin, or exhibits physical,psychological, or emotional symptoms of chronic stress. In certainembodiments, the skin rash and/or skin dehydration is associated withchronic stress and/or anxiety.

In another aspect, the invention provides methods of treating thepsychological, physical, or emotional changes associated with chronicstress comprising applying an effective amount of a dihydromyricetincompound, such as dihydromyricetin, to the individual's skin and/orscalp. In certain embodiments of the methods, the dihydromyricetincompound is provided in a hair product or a skin product described indetail elsewhere herein. In certain embodiments of the methods, thedihydromyricetin compound in the hair product or the skin product is ata concentration of about 0.01 μM to about 250 04. For example, thedihydromyricetin compound in the hair or skin product is at aconcentration of about 0.01 μM, about 0.05 μM, about 0.1 μM, about 0.2μM, about 0.3 μM, about 0.4 μM, 0.5 μM, about 0.6 μM, about 0.7 μM,about 0.8 μM, about 0.9 μM, about 1 μM, about 2 μM, about 3 μM, about 4μM, about 5 μM, about 6 μM, about 7 μM, about 8 μM, about 9 μM, about 10μM, about 25 μM, about 50 μM, about 100 μM, about 150 μM, about 200 μM,or about 250 μM, including any range in between these values. In certainembodiments of the methods, the dihydromyricetin compound in the hairproduct or the skin product is at a concentration of at least about 5%(w/v-%) or at least about 5% (w/w-%) to at least about 25% (w/v-%) or atleast about 25% (w/w-%). For example, the dihydromyricetin compound inthe hair or skin product is at a concentration of about at least about5% (w/v-%) or at least about 5% (w/w-%), at least about 6% (w/v-%) or atleast about 6% (w/w-%), at least about 7% (w/v-%) or at least about 7%(w/w-%), at least about 8% (w/v-%) or at least about 8% (w/w-%), atleast about 9% (w/v-%) or at least about 9% (w/w-%), at least about 10%(w/v-%) or at least about 10% (w/w-%), at least about 11% (w/v-%) or atleast about 11% (w/w-%), at least about 12% (w/v-%) or at least about12% (w/w-%), at least about 13% (w/v-%) or at least about 13% (w/w-%),at least about 14% (w/v-%) or at least about 14% (w/w-%), at least about15% (w/v-%) or at least about 15% (w/w-%), at least about 16% (w/v-%) orat least about 16% (w/w-%), at least about 17% (w/v-%) or at least about17% (w/w-%), at least about 18% (w/v-%) or at least about 18% (w/w-%),at least about 19% (w/v-%) or at least about 19% (w/w-%), at least about20% (w/v-%) or at least about 20% (w/w-%), at least about 21% (w/v-%) orat least about 21% (w/w-%), at least about 22% (w/v-%) or at least about22% (w/w-%), at least about 23% (w/v-%) or at least about 23% (w/w-%),at least about 24% (w/v-%) or at least about 24% (w/w-%), or at leastabout 25% (w/v-%) or at least about 25% (w/w-%), including any range inbetween these values.

In certain embodiments of the methods, the individual has alopecia,thinning hair, and/or baldness. In certain embodiments of the methods,the individual has telogen effluvium (TE) or alopecia areata. In certainembodiments of the methods, the individual exhibits signs of aging skin,e.g., the symptoms of aging skin described elsewhere herein. In certainembodiments, the individual may exhibit the physical, psychological,and/or emotional symptoms of chronic stress, including, but not limitedto, e.g., gingivitis, upset stomach or other digestive discomfort,dizziness and/or backache, insomnia, chest discomfort, moodiness,anxious, nervousness, aggression, intense mood swings, rash, poorconcentration, heightened confusion in stressful situations, inabilityto complete tasks, unstable blood pressure, hemorrhoids, varicose veins,panic attacks, and suicidal thoughts.

In certain embodiments of any methods described above, thedihydromyricetin compound is purified from whole plant or plant tissues.In certain embodiments, the dihydromyricetin compound is purified fromHovenia dulcis, Leptarrhena pyrolifolia, Pinus contorta, Ampelopsisgrossedentata, Glochidion sumatranum, Rhododendron ferrugineum, Ericaarborea, Salix hulteni, Manilkara zapota, Catharanthus roseus, Myricarubra, or Xantheoceras sorbifolia. In certain embodiments of any methodsdescribed above, the dihydromyricetin compound is synthesized usingmethods known in the art, and may be further purified. In certainembodiments, the dihydromyricetin compound is chemically synthesized. Incertain embodiments, the dihydromyricetin is purchased from adistributor.

Dihydromyricetin Compounds

As used herein, dihydromyricetin compounds include, without limitation,flavonoids having formula (I):

where each X is independently selected from hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, halo, alkoxyl, heteroalkyl,substituted heteroalkyl, heteroalkenyl, or substituted heteroalkenyl.

In certain embodiments of formula (I), X is independently selected from—CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —CH(CH₃)₂, —CH₂CH₂CH₂CH₃, —CH₂CH(CH₃)₂,—C(CH₃)₃, —CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₃, —C(CH₃)₂CH₂CH₃,—CH₂CH₂CH(CH₃)₂, —CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃,—CH₂CH₂C(CH₃)₂CH₃, —CH₂CH₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃,—CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₂CH₃, and—CH₂CH₂CH₂CH₂OCH₂CH₃.

In certain embodiments, the dihydromyricetin compound is of formula(II):

where each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene; and where each R is independently C₁₋₁₂ alkyl.

In certain embodiments, the dihydromyricetin compound is of formula(III):

where each Y is independently selected from is C₁₋₁₂ alkylene or C₁₋₁₂alkenylene; and where each R is independently C₁₋₁₂ alkyl.

In certain embodiments, the dihydromyricetin compound having formula (I)has hydrogen at position X. In such embodiments where X is a hydrogen,the dihydromyricetin compound is dihydromyricetin.

Dihydromyricetin compounds, such as dihydromyricetin, may be in the formof a salt which is produced by mixing dihydromyricetin with a base orthe like. The present disclosure provides for such salts ofdihydromyricetin compounds.

The terms “salts” as used herein refer to cosmetically acceptable orpharmaceutically acceptable base-addition salt forms of dihydromyricetincompounds, such as dihydromyricetin. Cosmetically acceptable salts arethose which are acceptable for topical applications and pharmaceuticallyacceptable salts are those which are acceptable for topicalpharmaceutical use, said salts being non-toxic. Said base-addition saltforms in particular are alkali metal, e.g. sodium or potassium, orammonium, or substituted ammonium salt forms, or salts with amino acidssuch as, for example, arginine, lysine and the like. Substitutedammonium as used herein refers to any non-toxic substituted ammonium ionknown or used in the art as a salt former and comprises mono-, di-, andin particular tri- or quaternary substituted ammonium salts, includingmono- or polycyclic systems. Substituents on ammonium for example arealkyl, cycloalkyl, alkenyl, substituted alkyl such as hydroxyalkyl,alkyloxyalkyl, (cycloalkyl)alkyl, arylalkyl such as benzyl, and thelike.

As used herein, dihydromyricetinR2R,3R)-3,5,7-trihydroxy-2-(3,4,5-trihydroxyphenyl)-2,3-dihydrochromen-4-one],also known as ampelopsin, is a flavonoid compound having the structuredepicted below:

Dihydromyricetin has been found in a variety of plants, including, e.g.,Hovenia dulcis (Ding et al. (1997) “Study on flavonoids in seeds ofHovenia dulcis.” Yao Xue Xue Bao. 32: 600-2), Leptarrhena pyrolifoliaand Pinus contorta (Stafford, et al. (1985) “Flavan-3-ol Biosynthesis:The Conversion of (+)-Dihydromyricetin to Its Flavan-3,4-Diol(Leucodelphinidin) and to (+)-Gallocatechin by Reductases Extracted fromTissue Cultures of Ginkgo biloba and Pseudotsuga menziesii.” PlantPhysiol. 78: 791-4), Ampelopsis grossedentata (Du et al. (2002)“Purification of (+)-dihydromyricetin from leaves extract of Ampelopsisgrossedentata using high-speed countercurrent chromatograph withscale-up triple columns.” J. Chromatogr. A. 973: 217-20), Glochidionsumatranum (Yin et al. (2010) “New galloylated flavanonols from theAustralian plant Glochidion sumatranum.” Planta Med. 76: 1877-1881),Rhododendron ferrugineum (Louis et al. (2010) “Phytochemicalcharacterization of Rhododendron ferrugineum and in vitro assessment ofan aqueous extract on cell toxicity.” Planta Med. 76: 1550-7), Ericaarborea (Nazimiyeh et al. (2008) “Antioxidant phenolic compounds fromthe leaves of Erica Arborea (Ericaceae).” Nat Prod Res. 22: 1385-92),Salix hulteni (Jeon et al. (2008) “Cytotoxic constituents from the barkof Salix hulteni.” Arch Pharm Res. 31: 978-982), Manilkara zapota (Ma etal. (2003) “Bioactive novel polyphenols from the fruit of Manilkarazapota (Sapodilla).” J Nat Prod. 66: 983-986), Catharanthus roseus(Cacace et al. (2003) “A flavonol O-methyltransderase from Catharanthusroseus performing two sequential methylations.” Phytochemistry. 62:127-137) and Xanthoceras sorbifolia (Ni et al. 2009) “Studies on thechemical constituents of Xanthoceras sorbifolia.” Zhong Yao Cai. 32:702-704). Other natural source of dihydromyricetin includes Myrica Rubra(also called Chinese bayberry, Japanese bayberry, red bayberry,yumberry, waxberry, or Chinese strawberry).

Dihydromyricetin can be extracted from these and other plants usingtechniques well known in the art. For example, dihydromyricetinextraction and purification methods are described in Yoo et al. (2006)“Recovery and pre-purification of (+)-dihydromyricetin from Hoveniadulcis.” Process Biochem 41: 567-570; Li et al. (2008) “Comparison ofrefluxing, ultrasonic- and microwave-assisted extraction ofdihydromyricetin from Ampelopsis grossedentata.” J AOAC Int. 91:1278-83; Du et al. (2002) “Purification of (+)-dihydromyricetin fromleaves extract of Ampelopsis grossedentata using high-speedcountercurrent chromatograph with scale-up triple columns.” J ChromatogA 973: 217-220; and others. Extraction methods described in Example 1may also be used,

The following method may also be used for extracting and purifyingdihydromyricetin from a whole plant or plant tissues:

Biomass extraction: The plant biomass is added to hot water at a ratioof 1:10 (w/v). The extraction begins with the biomass mixed with hotwater and is stirred at 100° C. for 6 h. The mixture is filtered throughfilter paper in a Buchner funnel under vacuum. These procedures arerepeated at least three times. Each water extract is collected, pooled,and concentrated at 40° C. under reduced pressure to decrease the volumeof the water extract to 20% of its original.

Liquid-liquid extraction: The concentrated water extract is added toorganic solvents (ethyl ether, chloroform, ethyl acetate, methyl-t-butylether, butanol) at a volume ratio of 4:1 for liquid-liquid extraction,and this is extracted at room temperature for 30 min. The extraction isrepeated at least three times, and the crude extracts were pooled anddried at room temperature under a reduced pressure.

Adsorbent treatment of the crude extract: The dried crude extract fromthe liquid-liquid extraction is dissolved in methanol at a ratio of 20(v/w) of methanol-to-dried crude extract, and several syntheticadsorbents were added and tested individually, including the activeclays P-1 and P-1G (Mizukalife Chemical Co., Japan), the activatedcarbons CA-1 and SX-PLUS (Norit, The Netherlands), and sylopute (FujiSilysia Chemical Ltd., Japan) at a ratio of 0.5 (w/w) of syntheticadsorbent to dried crude extract. The mixtures are stirred at roomtemperature for 30 min and filtered to obtain the filtration solution.The adsorbent cake thus obtained, is washed several times with ethylether/methanol (1:1, v/v) and the washings are combined with thefiltration solution. The solution is dried at 40° C. under a reducedpressure for the chromatography.

Silica gel low-pressure chromatography: The dried crude extract obtainedafter the adsorbent treatment is dissolved in methanol at a ratio of 10(v/w) of methanol-to-dried crude extract and then filtered throughdiatomaceous earth (Fuji Silysia Chemical Ltd., Japan). The filter aidis washed five times with methanol, and the washings are combined. Theresulting solution is applied to a 25 mm×400 mm column packed withsilica gel 60N (Merck, Germany), which is equilibrated with ethyl ether.The column is eluted using an isocratic method with the same solvent.The fractions containing (+)-dihydromyricetin are collected and dried byrotary evaporation.

The purity of dihydromyricetin extracted is assayed using methods knownin the art. For example, HPLC may be used to test the purity ofdihydromyricetin extracts. An HPLC system (Waters, Milford, USA) may beused for the analytical characterization of the intermediate and finalproducts. Purified dihydromyricetin extract may be analyzed using a C18column (4.6 mm×250 mm, 5 mm, Shiseido, Japan). The column is eluted witha water/acetonitrile gradient from 90:10 (v/v) to 30:70 (v/v) at a flowrate of 1.0 mL/min. The injection volume is 20 ml, and the effluent ismonitored at 254 nm with a UV detector. The dried residue is resolved inwater and used for the quantitative analysis of (+)-dihydromyricetin.Authentic (+)-dihydromyricetin (purity: 98%) may be purchased fromBiopurify Phytochemicals Ltd (China) and used as a standard.

Dihydromyricetin is also commercially available from a variety ofmanufacturers, including, e.g., Selleckchem, 2626 South Loop West, Suite225, Houston, Tex. 77054, USA; Shaanxi Huisheng Medicament TechnologyCo., Ltd., No. 78 Ping An Road, Yanglin High & New Technology Area,Shaanxi, China; Hunan 3W Botanical Extract Inc., C401, BLDG 1,International Enterprise Center, No. 188 Middle Huanbao Rd., Hunan,China; Phyto Nutraceutical Inc., TaiJia R,Wangcheng Xian, Changsha City,Hunan, China; and others.

Dihydromyricetin compounds can be produced by chemically alteringdihydromyricetin using methods well known to those of skill in the art.Alternatively, dihydromyricetin compounds can be chemically synthesizedusing methods well known to those of skill in the art.

C. Cosmetic Products Comprising Dihydromyricetin Compounds

In another aspect, the invention provides cosmetic products comprising adihydromyricetin compound, such as dihydromyricetin. In someembodiments, the dihydromyricetin compound in the cosmetic product is ata concentration of about 0.01 μM to about 250 04. In certainembodiments, a purified dihydromyricetin compound, such as purifieddihydromyricetin, is added to the cosmetic product or a material fromwhich the cosmetic product is made. In certain embodiments, the cosmeticproduct can be a hair product or a skin product, e.g., including, butnot limited to a hair or skin product described elsewhere herein.

In certain embodiments, the purified dihydromyricetin compound has beenadded directly to the cosmetic product or to a material from which thecosmetic product is made. A material used to make a cosmetic product cancomprise, e.g., a single ingredient or multiple ingredients, to whichfurther ingredients are added following the addition of thedihydromyricetin compound. The material can be a subset of ingredientsused to make the cosmetic product. The material can be treated (i.e.,heated, filtered, evaporated, mixed, liquefied, aerosolized, etc.)before or after the addition of the dihydromyricetin compound.

In certain embodiments, the dihydromyricetin compound is purified fromwhole plant or plant tissues (such as fruits). In certain embodiments,the dihydromyricetin compound is purified from Hovenia dulcis,Leptarrhena pyrolifolia, Pinus contorta, Ampelopsis grossedentata,Glochidion sumatranum, Rhododendron ferrugineum, Erica arborea, Salixhulteni, Manilkara zapota, Catharanthus roseus, Xantheoceras sorbifolia,or Myrica rubra. In certain embodiments, the dihydromyricetin compoundis chemically synthesized. In certain embodiments, the dihydromyricetincompound is purchased from a distributor.

In certain embodiments, the dihydromyricetin compound that has beenadded to the cosmetic product or to a material from which the cosmeticproduct is made is at least about 80% pure, at least about 85% pure, atleast about 90% pure, at least about 91% pure, at least about 92% pure,at least about 93% pure, at least about 94% pure, or at least about 95%pure. In certain embodiments, the dihydromyricetin compound that hasbeen added to the cosmetic product or to a material from which thecosmetic product is made is more than 95% pure, e.g., at least about 96%pure, at least about 97% pure, at least about 98% pure, at least about99% pure, or about 99% pure.

In certain embodiments, the dihydromyricetin compound in the cosmeticproduct (e.g., a hair product or skin product) is at a concentration ofabout 0.01 μM to about 250 04. For example, the dihydromyricetincompound in the cosmetic product is at a concentration of about 0.01 μM,about 0.05 μM, about 0.1 μM, about 0.2 μM, about 0.3 μM, about 0.4 μM,0.5 μM, about 0.6 μM, about 0.7 μM, about 0.8 μM, about 0.9 μM, about 1μM, about 2 μM, about 3 μM, about 4 μM, about 5 μM, about 6 μM, about 7μM, about 8 μM, about 9 μM, about 10 μM, about 25 μM, about 50 μM, about100 μM, about 150 μM, about 200 μM, or about 250 μM, including any rangein between these values. In certain embodiments, the dihydromyricetincompound in the cosmetic product (e.g., a hair product or skin product)is at a concentration of about at least about 5% (w/v-%) or at leastabout 5% (w/w-%), at least about 6% (w/v-%) or at least about 6%(w/w-%), at least about 7% (w/v-%) or at least about 7% (w/w-%), atleast about 8% (w/v-%) or at least about 8% (w/w-%), at least about 9%(w/v-%) or at least about 9% (w/w-%), at least about 10% (w/v-%) or atleast about 10% (w/w-%), at least about 11% (w/v-%) or at least about11% (w/w-%), at least about 12% (w/v-%) or at least about 12% (w/w-%),at least about 13% (w/v-%) or at least about 13% (w/w-%), at least about14% (w/v-%) or at least about 14% (w/w-%), at least about 15% (w/v-%) orat least about 15% (w/w-%), at least about 16% (w/v-%) or at least about16% (w/w-%), at least about 17% (w/v-%) or at least about 17% (w/w-%),at least about 18% (w/v-%) or at least about 18% (w/w-%), at least about19% (w/v-%) or at least about 19% (w/w-%), at least about 20% (w/v-%) orat least about 20% (w/w-%), at least about 21% (w/v-%) or at least about21% (w/w-%), at least about 22% (w/v-%) or at least about 22% (w/w-%),at least about 23% (w/v-%) or at least about 23% (w/w-%), at least about24% (w/v-%) or at least about 24% (w/w-%), or at least about 25% (w/v-%)or at least about 25% (w/w-%), including any range in between thesevalues.

In certain embodiments, the cosmetic products are hair products,including, but not limited to, e.g., shampoos, conditioners, masks,sprays or mists, gels, mousses, foams, serums, pastes, pomades, powders,oils, emulsions, creams, waxes, glazes, balms, tonics, lotions,ointments, polishes, lightening agents, straightening agents, relaxingagents, curling agents, or dyes. In certain embodiments, the cosmeticproducts are skin products, including, but not limited to, e.g.,lotions, moisturizers, facial polishers, facial cleaners or cleansers,sunscreens, skin patches, scrubs or exfoliating products, astringents,toners, masks, peels, gels, creams, balms, waxes, oils, salves, makeupremovers, insect repellents, soaps, makeup products (e.g., foundations,concealers or color correctors, blushers or rouges, lipsticks, lipglosses, lip balms, bronzers, setting sprays, powders, etc.), a mist, aspray, an ointment, a liniment, a topical analgesic, a topicalantihistamine, or an emulsion.

The invention also provide methods of applying a cosmetic productdescribed herein to a skin or hair area of an individual to treat hairloss, promote hair growth, delay or reverse signs of aging skin, ortreating chronic stress or anxiety.

D. Methods of Making Cosmetic Products Comprising DihydromyricetinCompounds

In yet another aspect, the invention provides methods of making acosmetic product (e.g., a hair product or a skin product) comprisingadding a dihydromyricetin compound, such as dihydromyricetin, to thecosmetic product or to a material from which the cosmetic product ismade. In some embodiments, a purified dihydromyricetin compound—such aspurified dihydromyricetin, is used in the methods, for example withpurity between at least about 80% pure and at least about 99% pure, orgreater than about 99% pure, as described herein. In certainembodiments, the methods are used to make a hair product or a skinproduct, as described elsewhere herein. In certain embodiments, thefinal concentration of the dihydromyricetin compound in the cosmeticproduct is between about 0.01 μM and about 250 μM.

In certain embodiments, the purified dihydromyricetin compound has beenadded directly to the cosmetic product or to a material from which thecosmetic product is made. A material used to make a cosmetic product cancomprise, e.g., a single ingredient or multiple ingredients, to whichfurther ingredients are added following the addition of thedihydromyricetin compound. The material can be a subset of ingredientsused to make the cosmetic product. The material can be treated (i.e.,heated, filtered, evaporated, mixed, liquefied, aerosolized, etc.)before or after the addition of the dihydromyricetin compound.

In certain embodiments, the dihydromyricetin compound is purified fromwhole plant or plant tissues. In certain embodiments, thedihydromyricetin compound is purified from Hovenia dulcis, Leptarrhenapyrolifolia, Pinus contorta, Ampelopsis grossedentata, Glochidionsumatranum, Rhododendron ferrugineum, Erica arborea, Salix hulteni,Manilkara zapota, Catharanthus roseus, Myrica rubra, or Xantheocerassorbifolia. In certain embodiments, the dihydromyricetin compound ischemically synthesized. In certain embodiments, the dihydromyricetincompound is purchased from a distributor.

In certain embodiments, the dihydromyricetin compound that has beenadded to the cosmetic product or to a material from which the cosmeticproduct is made is at least about 80% pure, at least about 85% pure, atleast about 90% pure, at least about 91% pure, at least about 92% pure,at least about 93% pure, at least about 94% pure, or at least about 95%pure. In certain embodiments, the dihydromyricetin compound that hasbeen added to the cosmetic product or to a material from which thecosmetic product is made is more than 95% pure, e.g., at least about 96%pure, at least about 97% pure, at least about 98% pure, at least about99% pure, or about 99% pure.

In certain embodiments of the methods, the dihydromyricetin compound inthe cosmetic product (e.g., a hair product or skin product) is at afinal concentration of about 0.01 μM, about 0.05 μM, about 0.1 μM, about0.2 μM, about 0.3 μM, about 0.4 μM, 0.5 μM, about 0.6 μM, about 0.7 μM,about 0.8 μM, about 0.9 μM, about 1 μM, about 2 μM, about 3 μM, about 4μM, about 5 μM, about 6 μM, about 7 μM, about 8 μM, about 9 μM, about 10μM, about 25 μM, about 50 μM, about 100 μM, about 150 μM, about 200 μM,or about 250 μM, including any range in between these values. In certainembodiments of the methods, the dihydromyricetin compound in thecosmetic product (e.g., a hair product or skin product) is at aconcentration of about at least about 5% (w/v-%) or at least about 5%(w/w-%), at least about 6% (w/v-%) or at least about 6% (w/w-%), atleast about 7% (w/v-%) or at least about 7% (w/w-%), at least about 8%(w/v-%) or at least about 8% (w/w-%), at least about 9% (w/v-%) or atleast about 9% (w/w-%), at least about 10% (w/v-%) or at least about 10%(w/w-%), at least about 11% (w/v-%) or at least about 11% (w/w-%), atleast about 12% (w/v-%) or at least about 12% (w/w-%), at least about13% (w/v-%) or at least about 13% (w/w-%), at least about 14% (w/v-%) orat least about 14% (w/w-%), at least about 15% (w/v-%) or at least about15% (w/w-%), at least about 16% (w/v-%) or at least about 16% (w/w-%),at least about 17% (w/v-%) or at least about 17% (w/w-%), at least about18% (w/v-%) or at least about 18% (w/w-%), at least about 19% (w/v-%) orat least about 19% (w/w-%), at least about 20% (w/v-%) or at least about20% (w/w-%), at least about 21% (w/v-%) or at least about 21% (w/w-%),at least about 22% (w/v-%) or at least about 22% (w/w-%), at least about23% (w/v-%) or at least about 23% (w/w-%), at least about 24% (w/v-%) orat least about 24% (w/w-%), or at least about 25% (w/v-%) or at leastabout 25% (w/w-%), including any range in between these values.

In certain embodiments, the cosmetic products are hair products,including, but not limited to, e.g., shampoos, conditioners, masks,sprays or mists, gels, mousses, foams, serums, pastes, pomades, powders,oils, emulsions, creams, waxes, glazes, balms, tonics, lotions,ointments, polishes, lightening agents, straightening agents, relaxingagents, curling agents, or dyes. In certain embodiments, the cosmeticproducts are skin products, including, but not limited to, e.g.,lotions, moisturizers, facial polishers, facial cleaners or cleansers,sunscreens, skin patches, scrubs or exfoliating products, astringents,toners, masks (e.g., facial masks), pads peels, gels, creams, balms,waxes, oils, salves, makeup removers, insect repellents, soaps, bathsalts, makeup products (e.g., foundations, concealers or colorcorrectors, blushers or rouges, lipsticks, lip glosses, lip balms,bronzers, setting sprays, powders, etc.), a mist, a spray, an ointment,a liniment, a topical analgesic, a topical antihistamine, an emulsion,or a facial treatment kits.

E. Kits and Articles of Manufacture

The invention also provides kits and articles of manufacture comprisinga cosmetic product of the invention packaged in a container with a labelindicating that the product is useful for promoting hair growth,reversing hair loss (or balding), promoting natural hair regrowth,increasing the thickness of thin (or thinning) hair, or preventing hairloss (or balding). The invention also provides kits and articles ofmanufacture comprising a cosmetic product of the invention packaged in acontainer with a label indicating that the product is useful fordelaying skin aging or reversing the signs of skin aging. In certainembodiments, the label may further indicate that the product has astress reducing, calming, and/or soothing effect. In certainembodiments, the label indicates the ingredients of the cosmetic productinclude a dihydromyricetin compound, such as dihydromyricetin.

EXAMPLES

The following examples are offered to illustrate, but not to limit theclaimed invention. It is understood that the examples and embodimentsdescribed herein are for illustrative purposes only and that variousmodifications or changes in light thereof will be suggested to personsskilled in the art and are to be included within the spirit and purviewof this application and scope of the appended claims.

Example 1 Use of Dihydromyricetin (DHM) in the Treatment ofStress-Induced Hair Loss

Dihydromyricetin (DHM) was extracted and purified from Hovenia dulcis.The extraction process involved the following steps. Hovenia dulcisplants (such as fruits) were extracted with 70% ethanol in 10-foldvolume of the plant tissues at 80-85° C. using soxhlet extraction(apparatus). The extraction was performed twice. The first extractionwas 2 hours, and the second extraction was 1.5 hours. The extract wascooled down to room temperature and filtered. The filtrate wasconcentrated to crystalize the compounds in the reactor and thencentrifuged to isolate precipitates. The precipitates were furtherextracted with 8-fold volume of 80% ethanol for 1 hour at 80-85° C.using soxhlet extraction (apparatus). The extract was cooled down toroom temperature and filtered. The filtrate was concentrated torecrystallize the compounds in the reactor. The filtrate was centrifugedto isolate precipitates, and the precipitates were then dried at 75-80°C. under ordinary atmospheric pressure. The dried material was groundinto powder which is the final product (BluCetin™). The productBluCetin™ contains≧98% DHM as tested by HPLC.

To determine whether topical application of DHM is toxic or causesallergic reaction, Vaseline mixed with 1 g DHM was applied to the shavedskin of c57BL/6J mice for six weeks. The animals did not exhibit rash,redness, or other responses associated with an allergic reaction. Inaddition, the animals did not exhibit any changes in body weight ormetabolic rate.

The effectiveness of DHM in the treatment of stress-induced hair losswas tested using two groups of eight week old C57BL/6J mice. Chronicstress was induced in the first group (n=20) by restraining each mousein the test group for 3 hours per day for 21 consecutive days in amodified 50 ml clear polystyrene conical centrifuge tube with multipleair holes for ventilation. The second group of mice (n=10) remainedundisturbed in their home cages (Fernandex-Vozmediano et al. (1994)“Contact dermatitis due to topical spironolactone.” Contact Dermatitis30: 118-9; Tresch, et al. (2011) “T cell-mediated acute localizedexanthematous pustulosis caused by finasteride.” J Allergy Clin Immunol129: 589-94). The completion of the 21 day stress-induction cycle wasfollowed by a two day withdrawal. The mice that were restrained inconical tubes exhibited diffuse hair loss, which was symptomatic ofchronic stress (FIG. 1). The rodents' stress levels were also assessedvia elevated plus maze (EPM) and open field test (OFT). EPM and OFT arebehavioral tests that are commonly used to assess anxiety by measuring amouse's locomotor activity and willingness to explore. When anxious,rodents naturally prefer enclosed dark spaces to opened brightly litspaces. In the EPM test, individual mice from each group were placed ona plus-shaped apparatus that was raised from the floor and had two openand two enclosed arms. Within the context of EPM, anxiety-relatedbehavior was measured by the degree to which a mouse avoided theunenclosed arms of the maze. As shown in FIG. 2A and FIG. 2B, theunstressed mice explored the open arms, whereas the chronically stressedmice avoided the open arms and restricted their movements to the closedarms. In OFT, each mouse was placed on an empty table surrounded bywalls, and the mouse's movements and the time it spent moving weremonitored. Within the context of OFT, anxiety-related behavior wasmeasured by the degree to which a mouse avoided the open center andstayed close to the walls. As shown in FIG. 2C and FIG. 2D, theunstressed mice explored center of the table, whereas the chronicallystressed mice avoided the center and remained in the corners. Takentogether, the results show that the chronically stressed mice wereobserved to exhibit both hair loss and behaviors associated withanxiety.

The chronically stressed mice were then divided into two groups: onegroup (n=10) was treatment with DHM (1 μM DHM in 1 oz. Vaseline) and theother group (n=10) was treated with Vaseline alone. Briefly, theVaseline (i.e., with or without 1 μM DHM) was applied to a mouse's baldspots once per day for six weeks. Following treatment, hair regrowth wasobserved in mice treated with DHM (FIG. 3). EPM and OFT were repeatedfor all mice (i.e., unstressed; stressed and treated with DHM; andstressed and treated with Vaseline alone). As shown in FIG. 4A and FIG.4B, the stressed mice treated with Vaseline+DHM were as willing toexplore the open arms of the EPM apparatus as the unstressed controlmice. In contrast, the stressed mice treated with Vaseline alonecontinued to avoid the open arms and confined their movements to theclosed arms. Similar behavior patterns were observed in OPT. Thestressed mice treated with Vaseline+DHM exhibited significantlyincreased locomotor activity and longer running distance comparable tothose of unstressed mice, indicating recovery from chronic stress (FIGS.4C and 4D). The stressed mice that were treated with Vaseline alonecontinued to exhibit anxiety-related behavior, such as the significantlack of motor activity (FIGS. 4C and 4D). Thus, both hair loss andbehavioral anxiety were observed to be reversed in DHM-treated stressedmice.

Example 2 Effects of Dihydromyricetin (DHM) Treatment on Hair Growth

A total of 40 seven-week-old, wild type C57BL/6J male mice were used forthe study. The mice were housed individually (one mouse/cage). The micewere randomly assigned to one of four treatment groups that contained 10mice per group. The treatment groups included a vehicle control, apositive control (minoxidil 2%), DHM 5% (w/w-%), and DHM 10% (w/w-%).

The vehicle used for all treatment groups was a mixture of alcoholdenat., propanediol, water, ethylhexylglycerin, and phenoxyethanol. TheDHM used in the DHM treatment groups has a purity of 98%.

The vehicle control contained the following percentage concentrations(w/w-%): 20% alcohol denat., 50% propanediol, 29.5% water, and 0.5%ethylhexylglycerin and phenoxyethanol.

The minoxidil 2% positive control contained the following percentageconcentrations (w/w-%): 20% alcohol denat., 2% minoxidil, 50%propanediol, 27.5% water, and 0.5% ethylhexylglycerin andphenoxyethanol.

The DHM 5% treatment contained the following percentage concentrations(w/w-%): 20% alcohol denat., 5% DHM, 50% propanediol, 24.5% water, and0.5% ethylhexylglycerin and phenoxyethanol.

The DHM 10% treatment contained the following percentage concentrations(w/w-%): 20% alcohol denat., 10% DHM, 50% propanediol, 19.5% water, and0.5% ethylhexylglycerin and phenoxyethanol.

The hair of the dorsal skin (2×5 cm) from every mouse was removed byclipping and topical application of calcium thioglycolate to depilatethe clipped area. The next day, mice without visible wounds were usedfor the study. The mice received a daily topical application of 1 ml oftheir respective treatment per day. Hair regrowth was quantified byanalyzing photographs that were taken of each mouse every 4 days aftertopical treatment for a total of 21 days (day 0, 4, 8, 12, 16, and 21).For each mouse, an area of 3 cm² of the back was analyzed. Allexperiments were continued for 21 days.

Evaluation the induction of anagen was performed on mice treated for 8days using a simple grading system: 0=no anagen; 1=<50% of the totalarea on anagen; 2=>50% and <100% on anagen; and 3=100% of the total areaon anagen. Evaluation of hair regrowth was performed on mice treated for12 days, using a simple grading system: 0=no hair; 1=<50% of the totalarea covered with hair; 2=>50% and <100% of the total area covered withhair; and 3=100% of the total area covered with hair.

To determine whether anagen induction was promoted by DHM, C57BL/6 micewere used, as the dorsal hair of C57BL/6 mice is known to have atime-synchronized hair growth cycle. The shaved skin of C57BL/6 mice ispink, which then darkens along with anagen initiation. After 8 days ofdepilation, an area of black skin was clearly visible in the mice.

As showed FIG. 5, treatment with DHM 10% was significantly different totreatment with the vehicle control, and similar to treatment with thepositive control (minoxidil 2%) in the induction of anagen. Treatmentwith DHM 10% also showed a statistically significant increase in hairregrowth as compared to treatment with the vehicle control (FIG. 6). Theresults of treatment with DHM 10% were also similar to the effectobserved with the minoxidil 2% positive control (FIG. 6).

1-28. (canceled)
 29. A cosmetic product comprising dihydromyricetincompound or a salt thereof, wherein the dihydromyricetin compound is offormula (I):

wherein each X is independently selected from hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, halo, alkoxyl,heteroalkyl, substituted heteroalkyl, heteroalkenyl, or substitutedheteroalkenyl, and wherein said dihydromyricetin compound in the productis at least about 0.01 μM.
 30. The cosmetic product of claim 29, whereinsaid dihydromyricetin compound in the product is about 0.01 μM to about250 μM.
 31. The cosmetic product of claim 29, wherein saiddihydromyricetin compound in the product is about 0.5 μM to about 10 μM.32. (canceled)
 33. The cosmetic product of claim 30, wherein saiddihydromyricetin compound is synthetic or purified from a whole plant ora plant tissue of Hovenia dulcis and added to the cosmetic product orthe material from which the cosmetic product is made.
 34. The cosmeticproduct of claim 30, wherein said dihydromyricetin compound is purifiedfrom a whole plant or a plant tissue of Leptarrhena pyrolifolia, Pinuscontorta, Ampelopsis grossedentata, Glochidion sumatranum, Rhododendronferrugineum, Erica arborea, Salix hulteni, Manilkara zapota,Catharanthus roseus, Myrica rubra, or Xanthoceras sorbifolia and addedto the cosmetic product or the material from which the cosmetic productis made.
 35. The cosmetic product of claim 34, wherein the purifieddihydromyricetin compound is at least about 95% pure.
 36. The cosmeticproduct of claim 30, wherein the cosmetic product is a hair product. 37.The cosmetic product of claim 36, wherein the hair product is a shampoo,a conditioner, or a hair spray.
 38. The cosmetic product of claim 30,wherein the cosmetic product is a skin product.
 39. The cosmetic productof claim 38, wherein the skin product is a moisturizer, a facialpolisher, a facial cleaner, a sunscreen, or a skin patch.
 40. Thecosmetic product of claim 30, wherein X is hydrogen.
 41. The cosmeticproduct of claim 30, wherein each X is independently selected from thegroup consisting of —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —CH(CH₃)₂, —CH₂CH₂CH₂CH₃,—CH₂CH(CH₃)₂, —C(CH₃)₃, —CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₃,—C(CH₃)₂CH₂CH₃, —CH₂CH₂CH(CH₃)₂, —CH₂CH₂CH₂CH₂CH₂CH₃, —CH₂C(CH₃)₂CH₂CH₃,—CH₂CH₂C(CH₃)₂CH₃, —CH₂CH₂CH₂CH(CH₃)₂, —CH₂CH(CH₂CH₃)CH₂CH₃,—CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₃, —CH₂CH₂CH₂OCH₂CH₃, and—CH₂CH₂CH₂CH₂OCH₂CH₃.
 42. The cosmetic product of claim 30, wherein thedihydromyricetin compound is of formula (II):

wherein each Y is independently selected from C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof.
 43. The cosmetic product of claim 30, wherein thedihydromyricetin compound is of formula (III):

wherein each Y is independently selected from C₁₋₁₂ alkylene or C₁₋₁₂alkenylene, and wherein each R is independently C₁₋₁₂ alkyl, or a saltthereof. 44-63. (canceled)
 64. The cosmetic product of claim 34, whereinthe cosmetic product is a hair product.